PrimeRoot: a cutting-edge technology designed to achieve precise and targeted large DNA insertion in plants

Genome editing techniques, especially clustered regularly interspaced short palindromic repeats (CRISPR), brought researchers into a new era of molecular plant breeding because it enabled them to make targeted modifications in plant genomes and transcriptomes. However, the successful incorporation of large DNA segments into plant genomes, necessary for high genetic gains and desired traits, remains a critical challenge. As there is an increasing demand for technologies that support chromosomal integration of large DNA inserts suitable for application in synthetic biology and plant breeding, PrimeRoot editors have presented a revolutionary solution to this issue, as they integrate enhanced prime editing guide RNA (PegRNA) designs, improved plant prime editor systems and advanced recombinases, helping in the precise insertion of DNA fragments of up to 11.1 kb into the plant genomes. Third-generation PrimeRoot editors further enhanced the precision and efficiency of transformation under different gene delivery systems. This technology holds enormous promise for accurately inserting long DNA sequences across different species of plants. This review highlights expected developments, opportunities, applications, advantages and challenges associated with PrimeRoot editors as well as the significance of expanding their applicability to more varieties of plants.